CHOLESTROL ESTIMATION

DR. BEENISH ZAKIINSTRUCTOR DEPARTMENT OF

BIOCHEMISTRY

OBJECTIVESSKILL:

At the end of the practical session, the student shall be able to demonstrate the ability to estimate serum Cholesterol through quantitative method.

KNOWLEDGE At the end of the session the student shall be

able to explain and interpret the principle and its clinical application.

Prerequisite For Test:

12 hours fasting from 8pm to 8 am.

Non-hemolyzed blood sample.

Method

This is an enzymatic colorimetric test using CHOD- PAP

method(Cholesterol Oxidase- Phenol 4-Aminoantipyrine

Peroxidase)

Principle

 Cholesterol esters+ H20 Cholesterol

esterase Cholesterol + fatty acid 

Cholesterol + H20 + 02 Cholesterol

oxidase Cholestenone + H202

H202 + 4 Aminoantipyrine + 3, 5- Dichlorophenol Peroxidase

Coloured quinonic derivative + 4 H20

Procedure:

Take 1 cuvette. Label as test (T).Mix well and let stand for 2

minutes at room temperature.Read optical density using green

filter. Working Reagent

Test

Sample 20 ulReagent 2000ul

CALCULATION

O.D of Test x 200 = Concentration of test

 O.D of standard

The unit is mg/dl

Normal Values of Serum Cholesterol

National Cholesterol Education Programme guidelines:

The Normal recommended level <170 mg/dlBorderline170-199 mg/dlHigh> 200 mg/dl

Cause of Hypercholesterolemia:

Increase in the amount of serum Cholesterol caused by because of:

Nephrotic SyndromeMyxodemaCoronary Artery Thrombosis

Causes of Hypocholesterolemia:

Decreased in amount of cholesterol can be due to:

HyperthyroidismMalabsorption syndromeHemolytic Jaundice

ANY QUESTIONS ?????

THANK YOU